Table 1

Success rates in protein expression, antibody isolation and immunohistochemistry

Step

Number

Percentage

(a) Mammalian expression of receptor extra-cellular domains

Sequenced confirmed clones

157

100%

Detected in supernatant by western blot

108

69%

50 μg purified product generated from 50 ml culture

67

62% (43% cumulative)

(b) Success rate of antigens in generation of antibodies

Mammalian antigens for selection

205

100%

Antigens generating antibody

181

88%

Bacterial antigens for selection

192

100%

Antigens generating antibody

103

54%

Total antigens for selection

404

100%

Antigens generating antibody

292

72%

(c) Success rate for primary antibody screening

Number of antibodies screened

38,164

100%

Number positive on primary screen

9,384

24.6%

Unique clones generated

7,236

77.1% (19% cumulative)

(d) Success rate for secondary antibody screening

Clones undergoing secondary specificity screening

4,437

100%

Positive in secondary screening

3,400

76.6%

Specificity demonstrated

2,725

80.1% (61% cumulative)

(e) Success rate in immunohistochemistry

Stained on tissue microarrays

381

100%

Positive staining observed

143

37%

Schofield et al. Genome Biology 2007 8:R254   doi:10.1186/gb-2007-8-11-r254